EP0521080B1 - Synergistic compositions - Google Patents
Synergistic compositions Download PDFInfo
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- EP0521080B1 EP0521080B1 EP91906872A EP91906872A EP0521080B1 EP 0521080 B1 EP0521080 B1 EP 0521080B1 EP 91906872 A EP91906872 A EP 91906872A EP 91906872 A EP91906872 A EP 91906872A EP 0521080 B1 EP0521080 B1 EP 0521080B1
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- Prior art keywords
- salt
- androst
- inhibitor
- diisopropylcarboxamide
- butylcarboxamide
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/565—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids not substituted in position 17 beta by a carbon atom, e.g. estrane, estradiol
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/02—Drugs for disorders of the urinary system of urine or of the urinary tract, e.g. urine acidifiers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
Definitions
- This invention comprises a composition containing one of certain selected compounds which are competitive inhibitors of steroid 5- ⁇ -reductase, combined with one of certain selected compounds which are uncompetitive inhibitors of steroid 5- ⁇ -reductase.
- the composition which contains this new combination of medicinal agents demonstrates improved suppression of plasma dihydrotestosterone (DHT) and improved prostate and seminal vesicle weight reduction.
- DHT dihydrotestosterone
- Steroid 5- ⁇ -reductase is a NADPH-dependent enzyme that mechanistically converts testosterone (T) to DHT.
- the individual compounds of the claimed compositions produce 5- ⁇ -reductase inhibiting activity by binding to a specific enzyme species at a specific stage of the reductive process.
- Combining inhibitors that bind to a different enzyme species at different stages of the reductive process has been found to lower the level of plasma DHT and to reduce the weight of the prostate and seminal vesicles to an extent beyond which a single inhibitor can achieve alone.
- Steroid 5- ⁇ -reductase is a NADPH-dependent enzyme that converts testosterone to DHT. Recognition of the importance of elevated DHT levels in various disease states has stimulated many efforts to synthesize inhibitors of this enzyme.
- the first inhibitor described was 4-androsten-3-one-17 ⁇ -carboxylic acid by Hsia and Voight in 1973. J. Invest. Dermat. 62 :224-227. (4R)-5,10-seco-19-norpregna-4,5-diene-3,10,20-triane was the next inhibitor to be described and also has found utility as an affinity label for 5- ⁇ -reductase. Robaire, B., et al., (1977), J. Steroid Biochem. 8 :307-310. (5 ⁇ ,20-R)-4-diazo-21-hydroxy-20-methylpregnan-3-one has been reported as a potent, time-dependent inhibitor of steroid 5- ⁇ -reductase.
- 17 ⁇ -acetoxy-6-methylene-pregn-4-ene-3,20-dione also has been shown to be a time-dependent inactivator of steroid 5- ⁇ -reductase.
- United States Patent 4,361,578 which issued June 2, 1986, describes a class of homosteroid enzyme inhibitors.
- United States Patent 4,191,759 discloses amides of 17 ⁇ -carboxy-4-androsten-3-one that are active as steroid 5- ⁇ -reductase inhibitors.
- Japanese Patents J60146855-A and J60116657-A disclose various aniline derivatives having numerous activities including 5- ⁇ -reductase inhibiting activity.
- Japanese Patent I60142941-A discloses phenyl-substituted ketones having 5- ⁇ -reductase inhibiting activity and European Patent EP173516-A discloses various phenyl-substituted amides having similar activity. Shiseido referenced terpene derivatives that are active inhibitors of steroid 5- ⁇ -reductase. Japanese Patent J59053417-A.
- enzyme forms to which such a steroidal-inhibitor might bind would include free enzyme, E, and enzyme binary complexes with cofactor, E-NADPH and E-NADP + .
- E free enzyme
- E-NADPH enzyme binary complexes with cofactor
- E-NADPH enzyme binary complexes with cofactor
- E-NADPH enzyme binary complexes with cofactor
- E-NADPH enzyme binary complexes with cofactor
- E-NADPH and E-NADP + enzyme binary complexes with cofactor
- the 4-aza steroidal inhibitors such as N-t-butyl-5- ⁇ -androst-1-ene-4-aza-17 ⁇ -carboxamide-3-one (compound A) (Liang et al., (1985), Endocrinology 117, 571-579)
- E-NADPH-1 enzyme-NADPH-1, Figure 1
- v is the observed velocity of product formation
- V m is the maximal enzyme velocity at saturating concentrations of the variable substrate (A)
- I is the concentration of the inhibitor with apparent inhibition constants of K is or K ii
- K a is the apparent Michaelis constant for the variable substrate.
- v V M A/[K a (1 + I/K is ) + A]
- v V M A/[K a + A(1 + I/K ii )]
- Velocities (v) determined at variable concentrations of substrate (A) and inhibitor (I) are evaluated by non-linear curve fittings with computer programs as described by Cleland (Cleland, W.W.
- Figure 4 represents a model incorporating conservation of drug substance.
- Curve A represents the uninhibited velocity
- curves B and C represent the presence of only competitive or uncompetitive inhibitors, respectively
- curve D presents conservation of inhibitor substance composed of half competitive and uncompetitive inhibitors.
- combination of inhibitors which function by the two different mechanisms is superior to the competitive inhibitors if the substrate concentration exceeds its K m , and is more efficient than the uncompetitive inhibitors at low substrate concentration below K m .
- a similar analysis versus the other substrate, NADPH would demonstrate no difference in inhibition efficiency since both kinetic models are described by equation 2.
- compositions comprising a 5 ⁇ -reductase inhibitor and an androgen blocking agent.
- a combination selected from “competitive” and “uncompetitive” inhibitors of 5- ⁇ -reductase are used in a pharmaceutical composition to affect the enzymatic reduction of testosterone to DHT.
- the “competitive” inhibiting components of this invention are compounds which form an enzyme-NADPH-inhibitor dead-end complex and the “uncompetitive” inhibiting components of this invention are compounds which form an enzyme-NADP + -inhibitor dead-end complex as described above.
- salts of these compounds containing a basic group are formed with organic or inorganic acids in the presence of a basic compound by methods known to the art.
- the compound is reacted with an inorganic or organic acid in an aqueous miscible solvent such as ethanol with isolation of the salt by removing the solvent or in an aqueous immiscible solvent when the acid is soluble therein, such as ethyl ether or chloroform, with the desired salt separating directly or isolated by removing the solvent.
- Exemplary of the acid addition salts which are included in this invention are maleate, fumarate, lactate, oxalate, methane-sulfonate, ethanesulfonate, benzenesulfonate, tartrate, citrate, hydrochloride, hydrobromide, sulfate, phosphate and nitrate salts.
- Pharmaceutically acceptable base addition salts of compounds of the invention containing an acidic group are prepared by known methods from organic and inorganic bases include nontoxic alkali metal and alkaline earth bases, for example, calcium, sodium, and potassium hydroxide; ammonium hydroxide, and nontoxic organic bases such as triethylamine, butylamine, piperazine, and (trihydroxymethyl)methylamine.
- Prodrug derivatives include O-esters, especially the tri-O-lower alkanoyl ester having from 2-8 carbon atoms in each alkanoyl group; O-methyl ethers or sulfate esters. Separated R and S stereoisomers are also useful.
- Compounds that are considered to be competitive steroid 5- ⁇ -reductase inhibitors as described above include: 3-oxa-4-aza steroidal compounds such as N-t-butyl-5- ⁇ -androst-1-ene-4-aza-17 ⁇ -carboxamide-3-one, (20R)-hydroxymethyl-4-methyl-4-aza-5-alpha-pregnane-3-one, 17 ⁇ -N,N-diisopropylcarboxamide-5- ⁇ -8(14)-androsten-4-methyl-4-aza-3-one, and 17 ⁇ -N-t-butylcarboxamide-5- ⁇ -8(14)-androsten-4-methyl-4-aza-3-one.
- 3-oxa-4-aza steroidal compounds such as N-t-butyl-5- ⁇ -androst-1-ene-4-aza-17 ⁇ -carboxamide-3-one, (20R)-hydroxymethyl-4-methyl-4-aza-5-alpha-pregnane-3-one, 17 ⁇ -N,N-diisopropyl
- 3-nitro steroidal compounds have been found to be competitive inhibitors including: 17 ⁇ -N,N-diisopropylcarboxamide-3-nitro-5- ⁇ -androst-3-ene, 17 ⁇ -N-t-butylcarboxamide-3-nitro-5- ⁇ -androst-3-ene, and 17 ⁇ -N,N-diisopropylcarboxamide-3-nitro-5 ⁇ -androst-2-ene.
- the claimed combinations inhibit steroid 5- ⁇ -reductase activity, they have therapeutic utility in treating diseases and conditions wherein decreases in DHT activity produce the desired therapeutic effect.
- diseases and conditions include acne vulgaris, seborrhea, female hirsutism, prostate diseases such as benign prostatic hypertrophy, and male pattern baldness.
- the ventral prostates When male rats are castrated and allowed to go untreated for 1 month, the ventral prostates involutes to approximately 10% of its original total cell number. If exogeneous testosterone is given to these castrated rats to restore the physiological serum level of testosterone, the involuted ventral prostate rapidly proliferates restoring its original cell content within 2-3 weeks. Using the ability to inhibit this testosterone induced regrowth of the rat ventral prostate, the efficacy and potency of twice daily oral administering of the "competitive" inhibitor and the "uncompetitive" inhibitor individually and in combination were compared.
- the competitive inhibitor being N-t-butyl-5- ⁇ -androst-1-ene-4-aza-17 ⁇ -carboxamide-3-one (Compound A) and the uncompetitive inhibitor being N-t-butyl-androst-3,5-diene-17 ⁇ -carboxamide-3-carboxylic acid (Compound C).
- the animals were administered the 5- ⁇ -reductase inhibiting compounds twice a day (BID) for 10 consecutive days.
- BID 5- ⁇ -reductase inhibiting compounds twice a day
- the test compound was dissolved in propylene glycol and diluted in normal saline.
- blood was collected from the animals and then they were sacrificed, the ventral prostrates were excised and weighed, and DHT levels were measured by the following procedure.
- Prostate tissue was excised, trimmed, weighed, minced and washed with phosphate buffer. The tissue then was homogenized in phosphate buffer and extracted by addition of ethyl acetate and mixing on an orbital mixer for forty-five minutes. The ethyl acetate was evaporated, the residue was reconstituted in ethanol, and was centrifuge filtered using 0.45 ⁇ M filter paper. The components then were separated using reverse-phase HPLC collecting the DHT fraction. The fraction was reduced to dryness and reconstituted in standard commercial DHT assay buffer. DHT levels then were measured using standard techniques such as radioimmunoassay.
- Solid or liquid pharmaceutical carriers are employed.
- Solid carriers include starch, lactose, calcium sulfate dihydrate, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid.
- Liquid carriers include syrup, peanut oil, olive oil, saline, and water.
- the carrier or diluent may include any prolonged release material, such as glyceryl monostearate or glyceryl distearate, alone or with a wax.
- the amount of solid carrier varies widely but, preferably, will be from about 25 mg to about 1 g per dosage unit.
- the preparation will be in the form of a syrup, elixir, emulsion, soft gelatin capsule, sterile injectable liquid such as an ampoule, or an aqueous or nonaqueous liquid suspension.
- the pharmaceutical preparations are made following conventional techniques of a pharmaceutical chemist involving mixing, granulating, and compressing, when necessary, for tablet forms, or mixing, filling and dissolving the ingredients, as appropriate, to give the desired oral or parenteral products.
- each active component of the claimed invention must be contemplated when formulating conventional dosage regimens.
- the individual compounds of the claimed combinations can be administered as a single pharmaceutical composition or consecutively in separate pharmaceutical compositions.
- One or more components can be incorporated into a timed release dosage unit form in which several doses are treated for delayed or sustained release of the medicament.
- Such dosage units may comprise sustained release granules, sugar centered spheres or multilayered tablets in each of which the availability of the active ingredient is controlled by coating with a lipid or polymeric material.
- Doses of the present combinations in a pharmaceutical dosage unit as described above will be an efficacious, nontoxic quantity selected from the range of 0.1 - 1000 mg/kg of each active compound, preferably 1-100 mg/kg.
- the selected dose is administered to a human patient in need of steroid 5- ⁇ -reductase inhibition from 1-6 times daily, topically, orally, rectally, by injection, or continuously by infusion.
- Oral dosage units for human administration preferably contain from 1 to 500 mg of active compound. Parenteral administration, which uses lower dosages is preferred. Oral administration, at higher dosages, however, also can be used when safe and convenient for the patient.
- the method of this invention of inhibiting steroid 5- ⁇ -reductase activity in mammals, including humans comprises administering internally to a subject in need of such inhibition an effective combination of steroid 5- ⁇ -reductase inhibiting compounds having both competitive and uncompetitive mechanisms of action.
- An oral dosage form for administering the claimed compounds and compositions is produced by screening, mixing and filling into hard gelatin capsules the ingredients in the proportions shown in Table II below.
- Table II Ingredients Amounts N-t-butyl-5- ⁇ -androst-1-ene-4-aza-17 ⁇ -carboxamide-3-one 100 mg N-t-butyl-androst-3,5-diene-17 ⁇ -carboxamide-3-carboxylic acid 100 mg Magnesium stearate 20 mg Lactose 300 mg
- sucrose, calcium sulfate dihydrate and claimed compounds and compositions shown in Table III below are mixed and granulated in the proportions shown with a 10% gelatin solution.
- the wet granules are screened, dried, mixed with the starch, talc and stearic acid, screened and compressed into a tablet.
- Table III Ingredients Amounts N-t-butyl-5- ⁇ -androst-1-ene-4-aza-17 ⁇ -carboxamide-3-one 100 mg N-t-butyl-androst-3,5-diene-17 ⁇ -carboxamide-3-carboxylic acid 100 mg Calcium sulfate dihydrate 300 mg Sucrose 40 mg Starch 20 mg Talc 10 mg Stearic acid 6 mg
- N-t-butyl-5- ⁇ -androst-1-ene-4-aza-17 ⁇ -carboxamide-3-one, 75 mg and N-t-butyl-androst-3,5-diene-17 ⁇ -carboxamide-3-carboxylic acid, 75 mg, are dispersed in 25 ml of normal saline to prepare an injectable preparation.
- the following compounds (expressed as base weight) are mixed together with 250 mg of lactose and 10 mg of magnesium stearate then filled into a hard gelatin capsule. These capsules are administered to a patient in need of steroid 5- ⁇ -reductase inhibiting activity from 1-6 times daily.
Abstract
Description
- This invention comprises a composition containing one of certain selected compounds which are competitive inhibitors of steroid 5-α-reductase, combined with one of certain selected compounds which are uncompetitive inhibitors of steroid 5-α-reductase. The composition which contains this new combination of medicinal agents demonstrates improved suppression of plasma dihydrotestosterone (DHT) and improved prostate and seminal vesicle weight reduction.
- Steroid 5-α-reductase is a NADPH-dependent enzyme that mechanistically converts testosterone (T) to DHT. The individual compounds of the claimed compositions produce 5-α-reductase inhibiting activity by binding to a specific enzyme species at a specific stage of the reductive process. Combining inhibitors that bind to a different enzyme species at different stages of the reductive process has been found to lower the level of plasma DHT and to reduce the weight of the prostate and seminal vesicles to an extent beyond which a single inhibitor can achieve alone.
- Steroid 5-α-reductase is a NADPH-dependent enzyme that converts testosterone to DHT. Recognition of the importance of elevated DHT levels in various disease states has stimulated many efforts to synthesize inhibitors of this enzyme.
- The first inhibitor described was 4-androsten-3-one-17β-carboxylic acid by Hsia and Voight in 1973. J. Invest. Dermat. 62:224-227. (4R)-5,10-seco-19-norpregna-4,5-diene-3,10,20-triane was the next inhibitor to be described and also has found utility as an affinity label for 5-α-reductase. Robaire, B., et al., (1977), J. Steroid Biochem. 8:307-310. (5α,20-R)-4-diazo-21-hydroxy-20-methylpregnan-3-one has been reported as a potent, time-dependent inhibitor of steroid 5-α-reductase. Blohm, T.R., et al., (1980), Biochem. Biophys. Res. Comm. 95:273-280; United States Patent 4,317,817, March 2, 1982. 17β-N,N-diethylcarbamoyl-4-methyl-4-aza-5-α-androstan-3-one is exemplary of a group of 4-aza steroid inhibitors of steroid 5-α-reductase described in United States Patent 4,377,584 which issued March 22, 1983, and in Liang, T., et al., (1983), J. Steroid Biochem. 19, 385-390. 17α-acetoxy-6-methylene-pregn-4-ene-3,20-dione also has been shown to be a time-dependent inactivator of steroid 5-α-reductase. Petrow, V., et al., (1981), Steroids 38:121-140.
- Other steroid 5-α-reductase inhibitors also have been described. United States Patent 4,361,578 which issued June 2, 1986, describes a class of homosteroid enzyme inhibitors. United States Patent 4,191,759 discloses amides of 17β-carboxy-4-androsten-3-one that are active as steroid 5-α-reductase inhibitors. Japanese Patents J60146855-A and J60116657-A disclose various aniline derivatives having numerous activities including 5-α-reductase inhibiting activity. Japanese Patent I60142941-A discloses phenyl-substituted ketones having 5-α-reductase inhibiting activity and European Patent EP173516-A discloses various phenyl-substituted amides having similar activity. Shiseido referenced terpene derivatives that are active inhibitors of steroid 5-α-reductase. Japanese Patent J59053417-A.
- Recently it has been shown that steroid 5-α-reductase follows an ordered kinetic mechanism (Houston et al., (1987), Steroids 49, 355-369; Metcalf, B., et al., (1989) Bioorganic Chemistry 17, 372-376) in which the nicotinamide cofactors are the first substrate binding to the enzyme and the last product released from the enzyme (Figure 1). Dead-end inhibitors of bisubstrate enzymes which follow ordered kinetic mechanisms can associate to one or more existing enzyme species. With steroid 5-α-reductase, enzyme forms to which such a steroidal-inhibitor might bind would include free enzyme, E, and enzyme binary complexes with cofactor, E-NADPH and E-NADP+. For example, the 4-aza steroidal inhibitors, such as N-t-butyl-5-α-androst-1-ene-4-aza-17β-carboxamide-3-one (compound A) (Liang et al., (1985), Endocrinology 117, 571-579), inhibit enzyme activity by forming an enzyme-NADPH-inhibitor dead-end complex (E-NADPH-1, Figure 1).
- In this context, the binding of inhibitors to each of these enzyme forms would be kinetically distinct as described in Irwin H. Segel, Enzyme Kinetics, Pub: John Wiley & Sons, Inc. (1975). The response of the velocity of enzyme catalysis with varying concentrations of testosterone in the presence of an inhibitor that binds to the E-NADPH complex can be described by equation 1; this model is denoted as competitive inhibition versus the variable substrate, in this case testosterone. Similarly, the model for an inhibitor that preferentially associates to the E-NADP+ complex would be described by equation 2; such a compound is referred to as an uncompetitive inhibitor versus the variable substrate. In equations 1 and 2, v is the observed velocity of product formation, Vm is the maximal enzyme velocity at saturating concentrations of the variable substrate (A), I is the concentration of the inhibitor with apparent inhibition constants of Kis or Kii, and Ka is the apparent Michaelis constant for the variable substrate.
- The effects of inhibitors which act by these two differing mechanisms under comparable conditions are shown in Figure 2; in this example, the concentration of each inhibitor has been set to be equal to its inhibition constant (Kii or Kis) and the Michaelis-constant for substrate (Km) has been set at 1 concentration unit. Curves B and C represent inhibitors that preferentially bind to the E-NADPH and E-NADP+ complexes while curve A represents the uninhibited velocity. Note that curves A and B intersect on the ordinate (competitive pattern) while curves A and C are parallel (uncompetitive pattern). From this plot, it can be seen that a competitive inhibitor (E-NADPH-I) is more efficient at low substrate concentration, while the uncompetitive inhibitor (E-NADP+-J) is more efficient at higher concentrations of substrate. The intersection point for these two curves occurs at the concentration of substrate that equals its Km.
- A single molecule that could bind equally well to both E-NADPH and E-NADP+ would demonstrate the additive effects in slope and intercept of the two double-reciprocal plots. This is described by equation 3, and is represented by curve D in Figure 2.
- Such additive inhibitory effects of a competitive inhibitor (N-t-butyl-5-α-androst-1-ene-4-aza-17β-carboxamide-3-one, compound A) and an uncompetitive inhibitor (N,N-diisopropyl-androst-3,5-diene-17β-carboxamide-3-carboxylic acid, compound B) have been demonstrated in vitro with double inhibition experiments; increasing concentrations of one inhibitor in the presence of the second induces greater enzyme inhibition (Figure 3), while also demonstrating mutually exclusive binding. For a given concentration of one of the inhibitors, which binds to E-NADPH (competitive) or E-NADP+ (uncompetitive), supplementation with any amount of the second would increase the observed enzyme inhibition over that of the reference inhibitor alone.
- In comparison, Figure 4 represents a model incorporating conservation of drug substance. Here, the calculated curves are based on the total drug substance relative to its respective inhibition constant:
- US 4,684,635 discloses compositions comprising a 5α-reductase inhibitor and an androgen blocking agent.
- No prior art of combining 5-α-reductase inhibitors of different mechanistic types into a pharmaceutical composition to achieve superior results is known to the applicants.
- A combination selected from "competitive" and "uncompetitive" inhibitors of 5-α-reductase are used in a pharmaceutical composition to affect the enzymatic reduction of testosterone to DHT. The "competitive" inhibiting components of this invention are compounds which form an enzyme-NADPH-inhibitor dead-end complex and the "uncompetitive" inhibiting components of this invention are compounds which form an enzyme-NADP+-inhibitor dead-end complex as described above.
- Also included are derivatives of these compounds which may either give rise to the parent compounds in vivo or be useful themselves, such as pharmaceutically acceptable addition salts. Salts of these compounds containing a basic group are formed with organic or inorganic acids in the presence of a basic compound by methods known to the art. For example, the compound is reacted with an inorganic or organic acid in an aqueous miscible solvent such as ethanol with isolation of the salt by removing the solvent or in an aqueous immiscible solvent when the acid is soluble therein, such as ethyl ether or chloroform, with the desired salt separating directly or isolated by removing the solvent. Exemplary of the acid addition salts which are included in this invention are maleate, fumarate, lactate, oxalate, methane-sulfonate, ethanesulfonate, benzenesulfonate, tartrate, citrate, hydrochloride, hydrobromide, sulfate, phosphate and nitrate salts. Pharmaceutically acceptable base addition salts of compounds of the invention containing an acidic group are prepared by known methods from organic and inorganic bases include nontoxic alkali metal and alkaline earth bases, for example, calcium, sodium, and potassium hydroxide; ammonium hydroxide, and nontoxic organic bases such as triethylamine, butylamine, piperazine, and (trihydroxymethyl)methylamine. Prodrug derivatives include O-esters, especially the tri-O-lower alkanoyl ester having from 2-8 carbon atoms in each alkanoyl group; O-methyl ethers or sulfate esters. Separated R and S stereoisomers are also useful.
- Compounds that are considered to be competitive steroid 5-α-reductase inhibitors as described above include: 3-oxa-4-aza steroidal compounds such as
N-t-butyl-5-α-androst-1-ene-4-aza-17β-carboxamide-3-one,
(20R)-hydroxymethyl-4-methyl-4-aza-5-alpha-pregnane-3-one,
17β-N,N-diisopropylcarboxamide-5-α-8(14)-androsten-4-methyl-4-aza-3-one, and
17β-N-t-butylcarboxamide-5-α-8(14)-androsten-4-methyl-4-aza-3-one.
Also 3-nitro steroidal compounds have been found to be competitive inhibitors including:
17β-N,N-diisopropylcarboxamide-3-nitro-5-α-androst-3-ene,
17β-N-t-butylcarboxamide-3-nitro-5-α-androst-3-ene, and
17β-N,N-diisopropylcarboxamide-3-nitro-5α-androst-2-ene. - Compounds that are considered to be uncompetitive steroid 5-α-reductase inhibitors as described above include:
N-t-butyl-androst-3,5-diene-17β-carboxamide-3-carboxylic acid or a salt thereof,
N,N-diisopropyl-androst-3,5-diene-17β-carboxamide-3-carboxylic acid or a salt thereof,
17β-(N,N-diisopropylcarboxamide)-estra-1,3,5(10)-triene-3-carboxylic acid or a salt thereof,
17β-(N-t-butylcarboxamide)-estra-1,3,5(10)-triene-3-carboxylic acid or a salt thereof,
17β-(N,N-diisopropylcarboxamide)-estra-1,3,5(10)-triene-3-sulfonic acid or a salt thereof,
20-α-(hydroxymethyl)-A-nor-5-α-pregn-1-ene-2-carboxylic acid or a salt thereof,
17β-(N-t-butylcarboxamide)-estra-1,3,5(10)-triene-3-sulfonic acid or a salt thereof,
17β-(N,N-diisorproyplcarboxamide)estra-1,3,5(10)-triene-3-phosphonic acid or a salt thereof,
17β-(N-t-butylcarboxamide)-estra-1,3,5(10)-triene-3-phosphonic acid or a salt thereof,
17β-(N,N-diisopropylcarboxamide)estra-1,3,5(10)-triene-3-phosphinic acid or a salt thereof,
17β-(N-t-butylcarboxamide)-estra-1,3,5(10)-triene-3-phosphinic acid or a salt thereof,
17β-N-t-butylcarboxamide-androst-3,5-diene-3-phosphinic acid or a salt thereof,
17β-N,N-diisopropylcarboxamide-androst-3,5-diene-3-phosphinic acid or a salt thereof,
17β-N-t-butylcarboxamide-androst-3,5-diene-3-phosphonic acid or a salt thereof, and
17β-N,N-diisopropylcarboxamide-androst-3,5-diene-3-phosphonic acid or a salt thereof. - Persons skilled in the art can readily determine if an inhibitor is a competitive or an uncompetitive type by the method set forth above.
- Because the claimed combinations inhibit steroid 5-α-reductase activity, they have therapeutic utility in treating diseases and conditions wherein decreases in DHT activity produce the desired therapeutic effect. Such diseases and conditions include acne vulgaris, seborrhea, female hirsutism, prostate diseases such as benign prostatic hypertrophy, and male pattern baldness.
- Certain combinations of the invention were tested for their in vivo potency in inhibiting steroid 5-α-reductase activity.
- When male rats are castrated and allowed to go untreated for 1 month, the ventral prostates involutes to approximately 10% of its original total cell number. If exogeneous testosterone is given to these castrated rats to restore the physiological serum level of testosterone, the involuted ventral prostate rapidly proliferates restoring its original cell content within 2-3 weeks. Using the ability to inhibit this testosterone induced regrowth of the rat ventral prostate, the efficacy and potency of twice daily oral administering of the "competitive" inhibitor and the "uncompetitive" inhibitor individually and in combination were compared.
- The competitive inhibitor being N-t-butyl-5-α-androst-1-ene-4-aza-17β-carboxamide-3-one (Compound A) and the uncompetitive inhibitor being N-t-butyl-androst-3,5-diene-17β-carboxamide-3-carboxylic acid (Compound C).
- To perform these experiments, a total of 50 male Sprague Dawley rats (from Harlan Sprague-Dawley, Inc.) of 250-300 gm starting weight were used. Forty-five of these animals were castrated and allowed to go untreated for 1 month to produce maximal ventral prostatic and seminal vesicle involution. After this 1 month period, 40 of the 45 castrated animals were implanted subcutaneously in the flank with a 2.5 cm long testosterone filled silastic capsule. The 50 animals were set up in 10 groups as follows:
- Group 1 -
- intact rats fed twice a day with vehicle alone (intact controls).
- Group 2 -
- castrated rats fed twice a day with vehicle and not implanted with testosterone capsule (castrate controls).
- Group 3 -
- castrated rats implanted with a testosterone capsule fed twice a day with vehicle (restoration control).
- Group 4 -
- castrated rats plus testosterone implant plus uncompetitive inhibitor (BID) 12.5 mg/kg.
- Group 5 -
- castrated rats plus testosterone implant plus uncompetitive inhibitor (BID) 25 mg/kg.
- Group 6 -
- castrated rats plus testosterone implant plus uncompetitive inhibitor (BID) 50 mg/kg.
- Group 7 -
- castrated rats plus testosterone implant plus competitive inhibitor (BID) 12.5 mg/kg.
- Group 8 -
- castrated rats plus testosterone implant plus competitive inhibitor (BID) 25 mg/kg.
- Group 9 -
- castrated rats plus testosterone implant plus competitive inhibitor (BID) 50 mg/kg.
- Group 10 -
- castrated rats plus testosterone implant plus uncompetitive inhibitor (BID) 50 mg/kg plus competitive inhibitor (BID) 50 mg/kg.
- The animals were administered the 5-α-reductase inhibiting compounds twice a day (BID) for 10 consecutive days. The test compound was dissolved in propylene glycol and diluted in normal saline. At the end of the treatment period blood was collected from the animals and then they were sacrificed, the ventral prostrates were excised and weighed, and DHT levels were measured by the following procedure.
- Prostate tissue was excised, trimmed, weighed, minced and washed with phosphate buffer. The tissue then was homogenized in phosphate buffer and extracted by addition of ethyl acetate and mixing on an orbital mixer for forty-five minutes. The ethyl acetate was evaporated, the residue was reconstituted in ethanol, and was centrifuge filtered using 0.45 µM filter paper. The components then were separated using reverse-phase HPLC collecting the DHT fraction. The fraction was reduced to dryness and reconstituted in standard commercial DHT assay buffer. DHT levels then were measured using standard techniques such as radioimmunoassay.
- In the rats treated with increasing amounts of a single inhibitor the realized prostate weight reduction plateaued at significantly higher levels than in the castrated control model, while in the combination-treated rats the prostate weights were reduced to castrate levels. This plateau dose-response effect regarding the competitive inhibitor compound A has also been reported in other studies. (Stoner, et al. (1987). Endocrinology, 120:774). Thus, the administration of the claimed combinations produces prostate weight reduction levels significantly lower than can be obtained by a single inhibitor alone.
- The claimed combinations are incorporated into convenient dosage forms such as capsules, tablets, or injectable preparations. Solid or liquid pharmaceutical carriers are employed. Solid carriers include starch, lactose, calcium sulfate dihydrate, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid. Liquid carriers include syrup, peanut oil, olive oil, saline, and water. Similarly, the carrier or diluent may include any prolonged release material, such as glyceryl monostearate or glyceryl distearate, alone or with a wax. The amount of solid carrier varies widely but, preferably, will be from about 25 mg to about 1 g per dosage unit. When a liquid carrier is used, the preparation will be in the form of a syrup, elixir, emulsion, soft gelatin capsule, sterile injectable liquid such as an ampoule, or an aqueous or nonaqueous liquid suspension.
- The pharmaceutical preparations are made following conventional techniques of a pharmaceutical chemist involving mixing, granulating, and compressing, when necessary, for tablet forms, or mixing, filling and dissolving the ingredients, as appropriate, to give the desired oral or parenteral products.
- The pharmacokinetic properties of each active component of the claimed invention must be contemplated when formulating conventional dosage regimens. To maximize its synergistic effect, the individual compounds of the claimed combinations can be administered as a single pharmaceutical composition or consecutively in separate pharmaceutical compositions. One or more components can be incorporated into a timed release dosage unit form in which several doses are treated for delayed or sustained release of the medicament. Such dosage units may comprise sustained release granules, sugar centered spheres or multilayered tablets in each of which the availability of the active ingredient is controlled by coating with a lipid or polymeric material.
- Doses of the present combinations in a pharmaceutical dosage unit as described above will be an efficacious, nontoxic quantity selected from the range of 0.1 - 1000 mg/kg of each active compound, preferably 1-100 mg/kg. The selected dose is administered to a human patient in need of steroid 5-α-reductase inhibition from 1-6 times daily, topically, orally, rectally, by injection, or continuously by infusion. Oral dosage units for human administration preferably contain from 1 to 500 mg of active compound. Parenteral administration, which uses lower dosages is preferred. Oral administration, at higher dosages, however, also can be used when safe and convenient for the patient.
- The method of this invention of inhibiting steroid 5-α-reductase activity in mammals, including humans, comprises administering internally to a subject in need of such inhibition an effective combination of steroid 5-α-reductase inhibiting compounds having both competitive and uncompetitive mechanisms of action.
- Following are the results of testing the combinations of this invention:
-
- The data in the above table demonstrate the synergistic effects of the "competitive" and "uncompetitive" steroid 5-α-reductase inhibitor combination on blocking testosterone induced regrowth of the involuted rat ventral prostate. Additionally, data was obtained showing that the testosterone levels were uneffected in all groups except in the castrate control.
- The following examples illustrate preparation of the claimed combinations and pharmaceutical compositions containing these compounds. The examples are not intended to limit the scope of the invention as defined hereinabove and as claimed below.
- An oral dosage form for administering the claimed compounds and compositions is produced by screening, mixing and filling into hard gelatin capsules the ingredients in the proportions shown in Table II below.
Table II Ingredients Amounts N-t-butyl-5-α-androst-1-ene-4-aza-17β-carboxamide-3-one 100 mg N-t-butyl-androst-3,5-diene-17β-carboxamide-3-carboxylic acid 100 mg Magnesium stearate 20 mg Lactose 300 mg - The sucrose, calcium sulfate dihydrate and claimed compounds and compositions shown in Table III below, are mixed and granulated in the proportions shown with a 10% gelatin solution. The wet granules are screened, dried, mixed with the starch, talc and stearic acid, screened and compressed into a tablet.
Table III Ingredients Amounts N-t-butyl-5-α-androst-1-ene-4-aza-17β-carboxamide-3-one 100 mg N-t-butyl-androst-3,5-diene-17β-carboxamide-3-carboxylic acid 100 mg Calcium sulfate dihydrate 300 mg Sucrose 40 mg Starch 20 mg Talc 10 mg Stearic acid 6 mg - N-t-butyl-5-α-androst-1-ene-4-aza-17β-carboxamide-3-one, 75 mg and N-t-butyl-androst-3,5-diene-17β-carboxamide-3-carboxylic acid, 75 mg, are dispersed in 25 ml of normal saline to prepare an injectable preparation.
- The following compounds (expressed as base weight) are mixed together with 250 mg of lactose and 10 mg of magnesium stearate then filled into a hard gelatin capsule. These capsules are administered to a patient in need of steroid 5-α-reductase inhibiting activity from 1-6 times daily.
- A. N-t-butyl-5-α-androst-1-ene-4-aza-17β-carboxamide-3-one, 100 mg; N-t-butyl-androst-3,5-diene-17β-carboxamide-3-carboxylic acid, 100 mg.
- B. N-t-butyl-5-α-androst-1-ene-4-aza-17β-carboxamide-3-one, 100 mg; 17β-(N-t-butylcarboxamide)-ester-1,3,5(10)-triene-3-carboxylic acid, 100 mg.
- C. 17β-N-t-butylcarboxamide-3-nitro-5-α-androst-3-ene, 100 mg; N-t-butyl-androst-3,5-diene-17β-carboxamide-3-carboxylic acid, 100 mg.
- D. 17β-N-t-butylcarboxamide-3-nitro-5-α-androst-3-ene, 100 mg; 17β-(N-t-butylcarboxamide)-ester-1,3,5(10)-triene-3-carboxylic acid, 100 mg.
Claims (8)
- A pharmaceutical composition comprising:(a) an uncompetitive inhibitor of steroid 5-α-reductase, and(b) a competitive inhibitor of steroid 5-α-reductase, and a pharmaceutically acceptable carrier.
- A composition according to claim 1 in which the competitive inhibitor is:
N-t-butyl-5-α-androst-1-ene-4-aza-17β-carboxamide-3-one;
17β-N,N-diisopropylcarboxamide-3-nitro-5-α-androst,-3-ene;
(2OR)-hydroxymethyl-4-methyl-4-aza-5-alpha-pregnane-3-one;
17β-N-t-butylcarboxamide-3-nitro-5-α-androst-3-ene;
17β-N,N-diisopropylcarboxamide-3-nitro-5-α-androst-2-ene;
17β-N,N-diisopropylcarboxamide-5-α-8(14)-androsten-4-methyl-4-aza-3-one; or
17β-N-t-butylcarboxamide-5-α-8(14)-androsten-4-methyl-4-aza-3-one. - A composition according to claim 1 in which the uncompetitive inhibitor is:
N-t-butyl-androst-3,5-diene-17β-carboxamide-3-carboxylic acid or a salt thereof;
N,N-diisopropyl-androst-3,5-diene-17β-carboxamide-3-carboxylic acid or a salt thereof;
17β-(N,N-diisopropylcarboxamide)-estra-1,3,5(10)-triene-3-carboxylic acid or a salt thereof;
17β-(N-t-butylcarboxamide)-estra-1,3,5(10)-triene-3-carboxylic acid or a salt thereof;
17β-(N,N-diisopropylcarboxamide)-estra-1,3,5(10)-triene-3-sulfonic acid or a salt thereof;
17β-(N-t-butylcarboxamide)-estra-1,3,5(10)-triene-3-sulfonic acid or a salt thereof;
17β-(N,N-diisopropylcarboxamide)-estra-1,3,5(10)-triene-3-phosphonic acid or a salt thereof;
17β-(N-t-butylcarboxamide)-estra-1,3,5(10)-triene-3-phosphonic acid or a salt thereof;
17β-(N,N-diisopropylcarboxamide)-estra-1,3,5(10)-triene-3-phosphinic acid or a salt thereof;
20-α-(hydroxymethyl)-A-nor-5-α-pregn-1-ene-2-carboxylic acid or a salt thereof;
17β-(N-t-butylcarboxamide)-estra-1,3,5(10)-triene-3-phosphinic acid or a salt thereof;
17β-N-t-butylcarboxamide-androst-3,5-diene-3-phosphinic acid or a salt thereof;
17β-(N-t-butylcarboxamide)-androst-3,5-diene-3-phosphonic acid or a salt thereof;
17β-N,N-diisopropylcarboxamide-androst-3,5-diene-3-phosphinic acid or a salt thereof; or
17β-N,N-diisopropylcarboxamide-androst-3,5-diene-3-phosphonic acid or a salt thereof. - A pharmaceutical composition according to claim 1 in which the competitive inhibitor is N-t-butyl-5-α-androst-1-ene-4-aza-17β-carboxamide-3-one, and the uncompetitive inhibitor is N-t-butyl-androst-3,5-diene-17β-carboxamide-3-carboxylic acid.
- A composition according to any one of claims 1 to 4 for use as an active therapeutic substance.
- A composition according to any one of claims 1 to 4 for use in inhibiting steroid 5-α-reductase activity.
- A composition according to any one of claims 1 to 4 for use in the treatment of acne vulgaris, seborrhea, female hirsuitism and prostate diseases such as benign prostatic hypertrophy and male pattern baldness.
- A process for preparing a pharmaceutical composition which process comprises bringing into association:(a) an uncompetitive inhibitor of steroid 5-α-reductase, and(b) a competitive inhibitor of steroid 5-α-reductase, and a pharmaceutically acceptable carrier or diluent.
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
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GB909006023A GB9006023D0 (en) | 1990-03-16 | 1990-03-16 | Synergistic compositions |
GB9006023 | 1990-03-16 | ||
PCT/US1991/001740 WO1991013550A1 (en) | 1990-03-16 | 1991-03-13 | Synergistic compositions |
Publications (3)
Publication Number | Publication Date |
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EP0521080A1 EP0521080A1 (en) | 1993-01-07 |
EP0521080A4 EP0521080A4 (en) | 1993-11-24 |
EP0521080B1 true EP0521080B1 (en) | 1996-12-11 |
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ID=10672772
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EP91906872A Expired - Lifetime EP0521080B1 (en) | 1990-03-16 | 1991-03-13 | Synergistic compositions |
Country Status (9)
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EP (1) | EP0521080B1 (en) |
JP (1) | JPH05506019A (en) |
AT (1) | ATE146080T1 (en) |
AU (1) | AU648551B2 (en) |
CA (1) | CA2076530A1 (en) |
DE (1) | DE69123573T2 (en) |
GB (1) | GB9006023D0 (en) |
HK (1) | HK1006542A1 (en) |
WO (1) | WO1991013550A1 (en) |
Families Citing this family (5)
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HUT71484A (en) * | 1992-05-20 | 1995-11-28 | Merck & Co Inc | Process for producing of 7beta-substituted-4-aza-5alpha-cholestan-ones as 5-alpha-reductase inhibitors and pharmaceuticals compositions containing them |
ATE227122T1 (en) * | 1993-09-03 | 2002-11-15 | Smithkline Beecham Corp | STABILIZED TABLET FORMULATION |
IL111467A0 (en) * | 1993-11-12 | 1994-12-29 | Merck & Co Inc | Pharmaceutical compositions comprising 7 beta -substituted -4-aza 5 alpha -cholestan-3-ones and 5 alpha reductase 1 inhibitors |
GB9411103D0 (en) * | 1994-06-03 | 1994-07-27 | Smithkline Beecham Corp | Compounds |
US5543417A (en) * | 1994-10-21 | 1996-08-06 | Merck & Co., Inc. | Combination method of treating acne using 4-AZA-5α-cholestan-ones and 4-AZA-5α-androstan-ones as selective 5α-reductase inhibitors with anti-bacterial, keratolytic, or anti-inflammatory agents |
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US4317817A (en) * | 1979-08-27 | 1982-03-02 | Richardson-Merrell Inc. | Novel steroid 5α-reductase inhibitors |
US4684635A (en) * | 1984-05-11 | 1987-08-04 | Norman Orentreich | Compositions and methods for inhibiting the action of androgens |
-
1990
- 1990-03-16 GB GB909006023A patent/GB9006023D0/en active Pending
-
1991
- 1991-03-13 JP JP91507241A patent/JPH05506019A/en active Pending
- 1991-03-13 WO PCT/US1991/001740 patent/WO1991013550A1/en active IP Right Grant
- 1991-03-13 EP EP91906872A patent/EP0521080B1/en not_active Expired - Lifetime
- 1991-03-13 CA CA002076530A patent/CA2076530A1/en not_active Abandoned
- 1991-03-13 AU AU75738/91A patent/AU648551B2/en not_active Ceased
- 1991-03-13 DE DE69123573T patent/DE69123573T2/en not_active Expired - Fee Related
- 1991-03-13 AT AT91906872T patent/ATE146080T1/en active
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1998
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Also Published As
Publication number | Publication date |
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JPH05506019A (en) | 1993-09-02 |
DE69123573T2 (en) | 1997-04-17 |
DE69123573D1 (en) | 1997-01-23 |
GB9006023D0 (en) | 1990-05-09 |
EP0521080A4 (en) | 1993-11-24 |
EP0521080A1 (en) | 1993-01-07 |
AU648551B2 (en) | 1994-04-28 |
HK1006542A1 (en) | 1999-03-05 |
AU7573891A (en) | 1991-10-10 |
CA2076530A1 (en) | 1991-09-17 |
ATE146080T1 (en) | 1996-12-15 |
WO1991013550A1 (en) | 1991-09-19 |
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